Cancer treatment using viruses and camptothecins

ABSTRACT

Mammalian subjects having a neoplasm are treated with a virus and a camptothecin compound, for example irinotecan or topotecan. The virus is selected from the group consisting of a Newcastle disease virus, a measles virus, a vesicular stomatitis virus, an influenza virus, a Sindbis virus, a picornavirus, and a myxoma virus. The treatment can also include administration of a monoclonal antibody against epidermal growth factor receptor, for example cetuximab.

BACKGROUND OF THE INVENTION

Coadministration of oncolytic viruses with other chemotherapeutic agentsis disclosed in WO 00/62735 (pages 35-36). See Kim D (Cancer Gene Ther2002; 9:959-960; Virotherapy for cancer: current status, hurdles andfuture directions) and Bell J C et al. (Cur Gene Ther 2002, 2:243-254;Oncolytic viruses: programmable tumour hunters) for recent reviews onanti-cancer virus therapy. Improvements in efficacy using such virustherapies are important to the field and getting approval and widespreaduse of the approach. Specifically, a drug which shows supra-additiveefficacy with a virus would be most advantageous.

The use of camptothecins as anticancer agents is reviewed inGarcia-Carbonero, et al., Clin. Cancer Res. (March 2002) 8: 641-661; andin Pizzolato J F and Saltz L B, The camptothecins. Lancet 2003361:2235-42. Camptothecins have antitumor activity based on theirbinding to and inhibition of topoisomerase I, a nuclear enzyme whichreduces torsional stress during DNA replication and which has animportant role in DNA replication. Topotecan and irinotecan are the twocamptothecins have been approved for clinical use by the US Food andDrug Administration (FDA). Other camptothecins are in development ascancer therapeutics (Ulukan and Swaan, (Campothecins: a review of theirchemotherapeutic potential. Drugs, 2002, 62:2039-57); andGarcia-Carbonero and Supko, 2002).

The treatment of cancers using certain mutant herpes viruses incombination with any of numerous anticancer agents, including irinotecanand topotecan, is disclosed in U.S. Patent Publication No. 2002/0071832(Fong, et al.), paragraphs 7 and 40. Methods of treating neoplasiasusing target cell-specific adenoviral vectors in combination withantineoplastic agents, including irinotecan or topotecan, are disclosedin U.S. Patent Publication No. 2003/0068307 (Yu, et al.) page 13. Seealso Nemunaitis, et al., Cancer Gene Ther. (2003) 10(5): 341-352; andMeck, et al., Cancer Res. (2001) 61(13): 5083-5089. Combined use ofirinotecan and cetuximab was approved in February 2004 by the U.S. FDAto treat colorectal cancer.

SUMMARY OF THE INVENTION

This invention provides a method for treating a mammalian subject havinga neoplasm, comprising administering to the subject a virus and acamptothecin compound in a combined amount effective to treat thesubject; wherein the virus is selected from the group consisting of aNewcastle disease virus, a measles virus, a vesicular stomatitis virus,an influenza virus, a Sindbis virus, a picornavirus, and a myxoma virus.In an embodiment of this invention the treatment further comprisesadministering to the subject a monoclonal antibody against epidermalgrowth factor receptor in an amount effective, in combination with thevirus and the camptothecin compound, to treat the subject.

This invention provides for the use of a virus and/or a camptothecincompound in the manufacture of a medicament for treating, in combinationwith the other ingredient mentioned, a subject having a neoplasm;wherein the virus is selected from the group consisting of a Newcastledisease virus, a measles virus, a vesicular stomatitis virus, aninfluenza virus, a Sindbis virus, a picornavirus, and a myxoma virus.This invention also provides the use of a monoclonal antibody againstepidermal growth factor receptor in the manufacture of a medicament fortreating, in combination with a virus as mentioned above and acamptothecin compound, a subject having a neoplasm.

This invention is based on the finding that anti-cancer viruses andcamptothecins in combination are effective against neoplastic cells. Toillustrate, a mesogenic strain of Newcastle disease virus andirinotecan, a camptothecin compound, have demonstrated a greater thanadditive level of in vivo antitumor activity, as shown in the examples.

DETAILED DESCRIPTION OF THE INVENTION

As used herein the transitional term “comprising” is open-ended. A claimutilizing this term can contain elements in addition to those recited insuch claim. Thus, for example, the claims can read on treatment regimensthat also include other therapeutic agents or therapeutic virus dosesnot specifically recited therein, as long as the recited elements ortheir equivalent are present.

As used herein “NDV” is an abbreviation for Newcastle Disease Virus. Asused herein “DLT” is an abbreviation for dose limiting toxicity. As usedherein the term “plaque-forming unit” (PFU) means one infectious virusparticle. As used herein “BPFU” means billion PFUs. As used herein “PP”means plaque-purified. Thus, for example PPMK107 means plaque-purifiedNewcastle Disease virus strain MK107. As used herein “PFU/m²”, which isa standard unit for expressing dosages, means PFUs per square meter ofpatient surface area. As used herein the term “replication-competent”virus refers to a virus that produces infectious progeny in cancercells.

In an embodiment of this invention the virus is replication-competent.

In accordance with this invention, when the virus is a Newcastle DiseaseVirus it can be of low (lentogenic), moderate (mesogenic) or high(velogenic) virulence. The level of virulence is determined inaccordance with the Mean Death Time in Eggs (MDT) test. (Alexander,“Chapter 27: Newcastle Disease” in Laboratory Manual for the Isolationand Identification of Avian Pathogens, 3^(rd) ed., Purchase, et al. eds.(Kendall/Hunt, Iowa), page 117.) Viruses are classified by the MDT testas lentogenic (MDT>90 hours); mesogenic (MDT from 60-90 hours); andvelogenic (MDT<60 hours). Mesogenic NDV is currently preferred.

In accordance with this invention, any conventional route or techniquefor administering viruses to a subject can be utilized. For examples ofroutes of administration refer to WO 00/62735. In one embodiment of thisinvention, the virus is administered systemically, for exampleintravenously. For intravenous administration of a therapeutic virus inaccordance with this invention, preferably the virus is a mesogenicstrain of Newcastle Disease Virus. In a preferred embodiment of thisinvention, from 12×10⁹ to 120×10⁹ PFU/m² per dose of a mesogenic strainof Newcastle Disease virus is administered intravenously to a humansubject, more preferably from 12×10⁹ to 48×10⁹ PFU/m² per dose. As usedherein “mg/m²” means milligrams per square meter of patient surfacearea.

In embodiments of this invention the picornavirus is a poliovirus, anechovirus, or a coxsackievirus. Examples of coxsackieviruses that aresuitable in accordance with this invention include the following types:A21, A13, A15 and A18. Examples of suitable echoviruses includeechovirus type 1.

As used herein the term “camptothecin compound” means that class ofcompounds considered to be camptothecins, camptothecin analogs,camptothecin derivatives or camptothecin conjugates. These compounds arebased on the characteristic five-ring backbone of camptothecin:

In accordance with this invention any camptothecin compound can beutilized. Examples of camptothecin compounds include irinotecan(CAMPTOSAR;7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin),topotecan (HYCAMPTIN;(S)-9-N,N-dimethylaminoethyl-10-hydroxycamptothecin),9-aminocamptothecin (9-amino-20(S)-camptothecin), 9-nitrocamptothecin(also called rubitecan), lurtotecan(7-(4-methylpiperazinomethylene)-10,11-ethylenedioxy-20(S)-camptothecin),exatecan, karenitecin, and a homocamptothecin. The structures andclinical information for some camptothecin compounds can be found inGarcia-Carbonero, et al., Clin. Cancer Res. (March 2002) 8: 641-661.Examples of camptothecin compounds can also be found in U.S. Pat. No.4,604,463, No. 6,403,569, and No. 5,004,758, and in WO 2004/012661, WO2003/101998, WO 2003/101996, WO 2003/101406, WO 2003/093274, WO2003/086471, WO 01/76597, WO 01/64194, WO 00/70275, WO 00/53607, WO99/17805, WO 99/17804, WO 99/05103, WO 98/35969, WO 97/28164, WO97/25332, WO 97/16454, the contents of all of which are incorporatedherein by reference.

In accordance with the combination therapy of this invention thecamptothecin compound can be administered from one month beforeadministration of the virus until one month after administration of thevirus. In more specific embodiments the camptothecin compound and thevirus are administered to the subject within a single twenty-four hourperiod; or the camptothecin compound is administered from twenty-fourhours to one month, preferably from twenty-four hours to one week,before administration of the virus; or the camptothecin compound isadministered to the subject from twenty-four hours to one month,preferably from twenty-four hours to one week, after administration ofthe virus. The dosing and administration techniques and schedules forcamptothecins and anti-cancer viruses are known in the art (See, e.g.Garcia-Carbonero, et al.; WO 00/62735; WO 2004/000209; and Pecora, etal., J. Clin. Oncol. (2002) 20(9): 2251-2266), and their optimizationfor a specific patient is within the ability of the skilled clinician.Irinotecan is usually administered to human patients in a dosage amountof from 62.5 to 125 mg/m² four times per week, or more preferably 80 to125 mg/m² four times per week; or from 300 to 350 mg/m² once every threeweeks, or more preferably 300 to 350 mg/m² once every three weeks.

In accordance with this invention any antibody against epidermal growthfactor receptor can be utilized. Chimeric and humanized monoclonalantibodies are preferred. Examples of suitable anti-EGF antibodiesinclude cetuximab (tradename: ERBITUX), ABX-EGF, MDX-447, h-R3, andEMD-7200 (see Mendelsohn J and Baselga J, “Status of epidermal growthfactor receptor antagonists in the biology and treatment of cancer”,2004 J Clin Oncol 21:2787-2799). Cetuximab is preferably administered tohuman patients intravenously, and is usually administered in an initialintravenous infusion of from 200 to 400 mg/m², followed approximatelyweekly thereafter by subsequent infusions of from 125 to 250 mg/m².

The subject that is treated in accordance with this invention can beeither a human subject or a non-human mammalian subject. In accordancewith this invention, any neoplasm can be treated, including but notlimited to the following: rectal cancer, pelvic cancer, colon cancer,lung cancer, breast cancer, prostate cancer, glioblastoma, renal cancer,pancreatic cancer, head and neck cancer, endometrial cancer,neuroblastoma, carcinoid, melanoma, ovarian cancer, sarcoma, cancer ofthe gastro-esophageal junction, gastric cancer, esophageal cancer, livercancer, and cervical cancer.

Although monitoring the treatment is not an essential aspect of theinvention, there are techniques for measuring the therapeutic effects ofthe treatment. These include, measuring the size of the tumor afteradministration of the virus, and a decrease in tumor size is a positiveresult.

The invention will be better understood by reference to the followingexamples, which illustrate but do not limit the invention describedherein. In the following examples 1 to 6, the NDV is a triple-plaquepurified MK107, which is an attenuated (mesogenic) version of NewcastleDisease Virus, described more fully in International Patent PublicationWO 00/62735, published October 26, 2000 (Pro-Virus, Inc.). The entirecontent of WO 00/62735 is hereby incorporated herein by reference.

EXAMPLES Example 1 NDV in Combination with Irinotecan

Athymic mice were injected subcutaneously with 10 million human HT1080fibrosarcoma cells. Five days later when the subcutaneous tumors wereapproximately 100 mm³ in size, groups of animals were treatedintraperitoneally with irinotecan (25 mg/kg) or vehicle. Two days lateranimals were treated intravenously with either NDV (6×10⁶ plaque formingunits, PFU) or vehicle. The incidence of complete tumor regression (CR,100% tumor reduction) was much higher in the group receiving bothirinotecan and NDV (60%) than either irinotecan alone (30%) or NDV alone(0%); see Table 1.

TABLE 1 Treatment of tumor-bearing mice with irinotecan 2 days beforetreatment with NDV yields greater complete tumor responses than eitheragent alone. Treatment Number of Mice CR, % Irinotecan 10 30% NDV 10  0%Both Irinotecan 10 60% and NDV Vehicle Control 10  0%

Example 2 NDV in Combination with Irinotecan

Athymic mice were injected subcutaneously with 10 million human HT1080fibrosarcoma cells. Seven days later when the subcutaneous tumors wereapproximately 125 mm³ in size, groups of animals were treatedintraperitoneally with irinotecan (25 mg/kg) or vehicle and thenapproximately one hour later they were treated intravenously with eitherNDV (6×10⁶ plaque forming units, PFU) or vehicle. The incidence ofcomplete tumor regression (CR, 100% tumor reduction) was much higher inthe group receiving both irinotecan and NDV (90%) than either irinotecanalone (50%) or NDV alone (0%), see Table 2.

TABLE 2 Treatment of tumor bearing mice with irinotecan the same day astreatment with NDV yields greater complete tumor responses than eitheragent alone. Treatment Number of Mice CR, % Irinotecan 10 50% NDV 10  0%Both Irinotecan 10 90% and NDV Vehicle Control 10  0%

Example 3 NDV in Combination with Irinotecan

Athymic mice were injected subcutaneously with 10 million human HT1080fibrosarcoma cells. Seven days later when the subcutaneous tumors wereapproximately 387 mm³ in size, groups of animals were intravenously witheither NDV (6×10⁶ plaque forming units, PFU) or vehicle. Two days later,the mice were then treated with treated intraperitoneally withirinotecan (25 mg/kg) or vehicle. The incidence of complete tumorregression (CR, 100% tumor reduction) was much higher in the groupreceiving both irinotecan and NDV (70%) than either irinotecan alone(10%) or NDV alone (0%), see Table 3.

TABLE 3 Treatment of tumor bearing mice with irinotecan two days aftertreatment with NDV yields greater complete tumor responses than eitheragent alone. Treatment Number of Mice CR, % Irinotecan 10 10% NDV 10  0%Both Irinotecan 10 70% and NDV Vehicle Control 10  0%

Example 4 NDV in Combination with Weekly Dosing of Irinotecan

Cancer patients are treated with NDV followed by treatment withirinotecan. In each 3 week portion of the 6 week cycle, NDV treatmentconsist of six total intravenous treatments given at three times perweek for two weeks followed by a one week rest period (see Table 4below). The first dose of each cycle consists of 12 to 24 billion PFU/m²(administered over 3 hours for course 1 and over 1 hour for all othercourses) followed by additional doses of between 24 to 48 billion PFU/m²(each dose administered over 1 hour). Irinotecan is given for fourconsecutive weeks on a weekly basis beginning during week 3 or 4 ofcycle 1 followed by two weeks without irinotecan therapy (As an example,see Table 4 below).Additional 6 week courses (also termed cycles) ofboth NDV and irinotecan are given to the patients.

TABLE 4 Combination of treatment of NDV using irinotecan (80 to 125mg/m²)given weekly x4. Cycles of treatment are repeated every 6 weeks.Cycle Week NDV? Irinotecan? 1 1 3 doses/wk for two No 2 weeks followedby 1 No 3 week off Yes, one dose over 90 minutes 4 3 doses/wk for twoYes, one dose over 90 minutes 5 weeks followed by 1 Yes, one dose over90 minutes 6 week off Yes, one dose over 90 minutes 2 1 3 doses/wk fortwo No 2 weeks followed by 1 No 3 week off Yes, one dose over 90 minutes4 3 doses/wk for two Yes, one dose over 90 minutes 5 weeks followed by 1Yes, one dose over 90 minutes 6 week off Yes, one dose over 90 minutes

Example 5 NDV in Combination Irinotecan Given Once Every 3 Weeks

Cancer patients are treated with NDV followed by treatment withirinotecan. NDV treatment consist of six total intravenous treatmentsgiven at three times per week for two weeks followed by a one week restperiod (see Table 5 below). The first dose of six consists of 12 to 24billion PFU/m² (administered over 3 hours for course 1 and over 1 hourfor all other courses) followed by a additional doses of 24 to 48billion PFU/m² (each dose administered over 1 hour). Patients begintheir irinotecan therapy during week 3 and are given one dose every 3weeks (See Table 5 below). Additional 3 week courses of both NDV andirinotecan are given to the patients.

TABLE 5 Combination of treatment of NDV using irinotecan given onceevery 3 wks. Cycles of treatment are repeated every 3 weeks. Cycle WeekNDV? Irinotecan? 1 1 3 doses/wk for two No 2 weeks followed by No 3 1week off Yes, one dose at 300 to 350 mg/m² over 30 minute intravenousinfusion 2 4 3 doses/wk for two No 5 weeks followed by No 6 1 week offYes, one dose at 300 to 350 mg/m² over 30 minute intravenous infusion

Example 6 NDV in Combination with Irinotecan and Cetuximab

Cancer patients are treated with both NDV and irinotecan as in examples4 and 5, except that they additionally receive treatment with cetuximab[ERBITUX, a monoclonal antibody (mAb) against the epidermal growthfactor receptor (EGFR)]. Cetuximab dosing begins on week 3 or week 4.The cetuximab dose is 200 to 400 mg/m² for the first intravenous (IV)infusion [administered as a 120 minute IV infusion (with a maximalinfusion rate of 5 mL/min)] then 125 to 250 mg/m² [infused IV over 60minutes] administered weekly thereafter. Some patients may also receivean initial test dose of cetuximab of 20 mg. Diphendydramine (50 mg IV)is commonly given to help lessen any infusion reactions due tocetuximab.

What is claimed is:
 1. A method of treating a mammalian subject having a neoplasm, comprising administering to the subject a Newcastle disease_virus and a camptothecin compound in a combined amount effective to treat the subject.
 2. The method of claim 1, further comprising administering to the subject a monoclonal antibody against epidermal growth factor receptor in an amount effective, in combination with the virus and the camptothecin compound, to treat the subject.
 3. The method of claim 2, wherein the monoclonal antibody is cetuximab.
 4. The method of claim 1, wherein the virus is replication-competent.
 5. The method of claim 1, wherein the virus is a mesogenic strain of Newcastle disease virus
 6. The method of claim 1, wherein the virus is administered intravenously.
 7. The method of claim 1 wherein the camptothecin compound is selected from the group consisting of topotecan, 9-aminocamptothecin, exatecan, karenitecin, rubitecan, lurtotecan, and a homocamptothecin.
 8. The method of claim 1, wherein the camptothecin compound is irinotecan.
 9. The method of claim 1, wherein the camptothecin compound and the virus are administered to the subject within a single twenty-four hour period.
 10. The method of claim 1, wherein the camptothecin compound is administered from twenty-four hours to one month before administration of the virus.
 11. The method of claim 10, wherein the camptothecin compound is administered from twenty-four hours to one week before administration of the virus.
 12. The method of claim 1, wherein the camptothecin compound is administered to the subject from twenty-four hours to one month after administration of the virus.
 13. The method of claim 12, wherein the camptothecin compound is administered to the subject from twenty-four hours to one week after administration of the virus. 